Method for treating ovarian cancer and/or hodgkin lymphomia with a gold(iii) complex

ABSTRACT

A method for treating cancer and gold(III) complexes with diaminocyclohexane ligand as anticancer agents. Also described are a pharmaceutical composition incorporating the gold(III) complexes and a method of synthesizing the gold(III) complexes.

RELATED APPLICATIONS

This application claims the priority of the filing date of the U.S.Provisional Patent Application No. 62/262,550 filed Dec. 3, 2015, thedisclosure of which is hereby incorporated herein by reference in itsentirety.

STATEMENT OF FUNDING ACKNOWLEDGEMENT

This project was funded by the National Plan for Science and Innovation(MARIFAH)-King Abdulaziz City for Science and Technology (KACST) throughthe Science and Technology Unit at King Fahd University of Petroleum andMinerals (KFUPM) of Saudi Arabia, award No. 14-MED64-04.

BACKGROUND OF THE DISCLOSURE

Technical Field

The present disclosure relates to gold(III) complexes with anti-canceractivity and a method of treating cancer.

Description of the Related Art

The “background” description provided herein is for the purpose ofgenerally presenting the context of the disclosure. Work of thepresently named inventors, to the extent it is described in thisbackground section, as well as aspects of the description which may nototherwise qualify as prior art at the time of filing, are neitherexpressly or impliedly admitted as prior art against the presentinvention.

One of the most important areas of study in contemporary bioinorganicchemistry is the development of new metal-based drugs (S. L. Best and P.J. Sadler, Gold Bull., vol. 29, no. 3, pp. 87-93, 1996; A. Casini, G.Kelter, C. Gabbiani, M. A. Cinellu, G. Minghetti, D. Fregona, H.-H.Fiebig, and L. Messori, J. Bio. Inorg. Chem., vol. 14, no. 7, pp.1139-1149, 2009; S. H. van Rijt and P. J. Sadler, Drug Discov. Today,vol. 14, no. 23-24, pp. 1089-97, 2009; X. Wang and Z. Guo, DaltonTrans., no. 12, pp. 1521-32, 2008; C. Frank Shaw III, Chem. Rev., vol.99, no. I, pp. 2589-2600, 1999; P. J. Sadler and R. E. Sue, Met. BasedDrugs, vol. 1, pp. 107-44, 1994; I. Ott, Coord. Chem. Rev., vol. 253,no. 11-12, pp. 1670-1681, 2009; A. C. and L. M. Chiara Gabbiani, GoldBull, vol. 40, pp. 73-81, 2007; M. A. Cinellu, L. Maiore, M. Manassero,A. Casini, M. Arca, H.-H. Fiebig, G. Kelter, E. Michelucci, G.Pieraccini, C. Gabbiani, and L. Messori, ACS Med. Chem. Lett., vol. 1,no. 7, pp. 336-339, 2010; and A. Bindoli, M. P. Rigobello, G. Scutari,C. Gabbiani, A. Casini, and L. Messori, Coord. Chem. Rev., vol. 253, no.11-12, pp. 1692-1707, 2009, each incorporated herein by reference intheir entirety). Despite the successful use of cisplatin as achemotherapy agent, side effects and resistance have been observed (A.Casini, C. Hartinger, C. Gabbiani, E. Mini, P. J. Dyson, B. K. Keppler,and L. Messori, J. Inorg. Biochem., vol. 102, no. 3, pp. 564-575, 2008;and V. Milacic, D. Fregona, and Q. P. Dou, Histol. Histopathol., vol.23, no. 1, pp. 101-108, 2008, each incorporated herein by reference intheir entirety). The use of gold in anti-rheumatic treatment supportsits pharmaceutical importance (M. W. Whitehouse, Inflammopharmacology,vol. 16, no. 3, pp. 107-9, 2008; G. G. Graham, M. W. Whitehouse, and G.R. Bushell, Inflammopharmacology, vol. 16, no. 3, pp. 126-32, 2008; A.Casini, M. A. Cinellu, G. Minghetti, C. Gabbiani, M. Coronnello, E.Mini, and L. Messori, J. Med. Chem., vol. 49, no. 18, pp. 5524-31, 2006;and L. Ronconi, L. Giovagnini, C. Marzano, F. Betti, R. Graziani, G.Pilloni, and D. Fregona, Inorg. Chem., vol. 44, no. 6, pp. 1867-81,2005, each incorporated herein by reference in their entirety). Recentstudies have shown that several gold(III) complexes are highly cytotoxicagainst different tumor cells (L. Giovagnini, L. Ronconi, D. Aldinucci,D. Lorenzon, S. Sitran, and D. Fregona, J. Med. Chem., vol. 48, no. 5,pp. 1588-95, 2005; A. Casini, M. A. Cinellu, G. Minghetti, C. Gabbiani,M. Coronnello, E. Mini, and L. Messori, J. Med. Chem., vol. 49, no. 18,pp. 5524-5531, 2006; and D. Saggioro, M. P. Rigobello, L. Paloschi, A.Folda, S. A Moggach, S. Parsons, L. Ronconi, D. Fregona, and A. Bindoli,Chem. Biol., vol. 14, no. 10, pp. 1128-39, 2007, S. S. Al-Jaroudi, M.Monim-ul-Mehboob, M. Altaf, M. Fettouhi, M. I. M. Wazeer, S. Altuwaijri,and A. A. Isab, New J. Chem., vol 38, pp. 3199-3211, 2014; M.Arsenijevic, M. Milovanovic, V. Volarevic, A. Djekovic, T. Kanjevac, N.Arsenijevic, S. Dukic, Z. D. Bugarcic, Med. Chem., vol. 8, no. 1, pp.2-8, 2012, each incorporated herein by reference in their entirety).Oxaliplatin, a platinum-based drug, is coordinated to(1R,2R)-(−)-1,2-diaminocyclohexane and a labile oxalate ligand (G. Sava,A. Bergamo, and P. J. Dyson, Dalton Trans., vol. 40, no. 36, pp.9069-75, 2011; and L. Kelland, Nat. Rev. Cancer, vol. 7, no. 8, pp.573-84, 2007, each incorporated herein by reference in their entirety).

Interactions of gold(III) complexes with biomolecules, such assulfur-containing amino acids, thiols, or thioethers, are thought to bethe cause of the cytotoxic effect of gold(III) complexes (A. V.Vuja{hacek over (c)}ić, J. Z. Savić, S. P. Sovilj, K. MészárosSzécsényi, N. Todorovic, M. {hacek over (Z)}. Petković, and V. M. Vasić,Polyhedron, vol. 28, no. 3, pp. 593-599, 2009, incorporated herein byreference in its entirety). The toxic effects arise due to thecoordination of the thiol and thioether groups of the side chains inproteins, peptides and amino acids followed by the reduction ofgold(III) to gold(I) and subsequently to toxic gold(0) (T. Kolev, B. B.Koleva, S. Y. Zareva, and M. Spiteller, Inorg. Chim. Acta, vol. 359, no.13, pp. 4367-4376, 2006; J. Zou, Z. Guo, J. A. Parkinson, Y. Chen, andP. J. Sadler, Chem. Commun., vol. 8, pp. 1359-1360, 1999; J. A.Cuadrado, W. Zhang, W. Hang, and V. Majidi, J. Env. Monit, vol. 2, no.4, pp. 355-359, 2000; and P. L. Witkiewicz. and C. F. Shaw. III, J.Chem. Soc. Chem. commun, pp. 1111-1114, 1981, each incorporated hereinby reference in their entirety). The electrochemistry of the interactionof guanosine-5-phosphate with gold(III) ethylenediamine complexes wasstudied by Zhu et al. (S. Zhu, W. Gorski, D. R. Powell, and J. A.Walmsley, Inorg. Chem., vol. 45, no. 6, pp. 2688-2694, 2006,incorporated herein by reference in its entirety). The interaction ofthe dipeptide Gly-Met with gold(III) ethylenediamine showed theformation of Gly-Met sulfoxide along with a free ethylenediamine ligandthrough a two-step decomposition reaction (B. D. Gli{hacek over (s)}ić,M. I. Djuran, Z. D. Stanić, and S. Rajković, Gold Bull, vol. 47, no.1-2, pp. 33-40, 2014, incorporated herein by reference in its entirety).The substitution and reduction steps for the reaction of some gold(III)complexes with the biological thiols, L-cysteine, L-methionine, andglutathione were studied in detail by Durović et al. (M. D. Durović, Z.D. Bugar{hacek over (c)}ić, F. W. Heinemann, and R. van Eldik, Dalton.trans., vol. 43, no. 10, pp. 3911-21, 2014, incorporated herein byreference in its entirety). The interaction between [Au(CN)₄]⁻ andglutathione (GSH) at pH 7.4 showed the reduction of gold(III) to gold(I)along with the dimerization of GSH into its disulfide (GSSG²⁻)derivative as the reaction product through a two-step decompositionreaction (P. M. Yangyuoru, J. W. Webb, and C. F. Shaw, J. Inorg.Biochem., vol. 102, no. 3, pp. 584-93, 2008; and B. A. Al-Maythalony, M.I. M. Wazeer, and A. A. Isab, Inorg. Chim. Acta, vol. 363, no. 13, pp.3244-3253, 2010, each incorporated herein by reference in theirentirety).

Therefore, it is an objective of this disclosure to provide gold(III)complexes with anti-cancer activity and a method for treating cancer.

BRIEF SUMMARY

The foregoing paragraphs have been provided by way of generalintroduction, and are not intended to limit the scope of the followingclaims. The described embodiments, together with further advantages,will be best understood by reference to the following detaileddescription taken in conjunction with the accompanying drawings.

The first aspect of the disclosure relates to a method for treatingcancer, comprising administering an effective amount of at least one ofa gold(III) complex represented by formula (I), a gold(III) complexrepresented by formula (II):

a pharmaceutically acceptable salt, solvate, prodrug, or a combinationthereof to a subject;

where the cancer is ovarian cancer, Hodgkin lymphoma, or both;

each of R₁ is independently a hydrogen, an optionally substituted C₁-C₈alkyl group, or an optionally substituted C₆-C₈ aryl group;

each of R₂ is independently a hydrogen, a halogen, a hydroxyl, an amino,a nitro, a cyano, an optionally substituted alkyl, an optionallysubstituted cycloalkyl, an optionally substituted heterocyclyl, anoptionally substituted arylalkyl, an optionally substituted heteroaryl,an optionally substituted alkoxyl, an optionally substituted aryl, anoptionally substituted alkenyl, a N-monosubstituted amino group, or aN,N-disubstituted amino group; and

X is at least one pharmaceutically acceptable anion, such as chloride,bromide, and iodide.

In one embodiment, R₁ is hydrogen.

In one embodiment, R₂ is hydrogen.

In one embodiment, X is chloride.

In one embodiment, the gold(III) complex represented by formula (I) is

and the gold(III) complex represented by formula (II) is

In one embodiment, the gold(III) complex represented by formula (I) orthe gold(III) complex represented by formula (II) is administered as asalt having at least one counterion which is at least onepharmaceutically acceptable anion selected from the group consisting offluoride, chloride, bromide, iodide, nitrate, sulfate, phosphate,methanesulfonate, ethanesulfonate, p-toluenesulfonate, salicylate,malate, maleate, succinate, tartrate, citrate, acetate, perchlorate,trifluoromethanesulfonate, acetylacetonate, hexafluorophosphate, andhexafluoroacetylacetonate.

In one embodiment, the at least one counterion is chloride.

In one embodiment, the ovarian cancer is resistant to cisplatin.

In one embodiment, the Hodgkin lymphoma is classical Hodgkin lymphoma.

In one embodiment, the method further comprises measuring aconcentration of a biomarker and/or detecting a mutation in thebiomarker before and/or after the administering.

In one embodiment, the biomarker is at least one selected from the groupconsisting of BRCA1, BRCA2, CCL17, CD163, CD30, NF-κB, Gal-1, CA125,HE4, mesothelin, transthyretin, ApoA1, VCAM, IL-6, IL-8, B7-H4, serumamyloid A, transferrin, osteopontin, kallikreins, OVX1, VEGF, AGR-2,inhibin, M-CSF, uPAR, EGF receptor, lysophosphatidyl acid,beta2-microglobulin, miRNA, and Epstein-Barr virus DNA.

In one embodiment, the concentration of the biomarker is measured withan ELISA assay and/or the mutation in the biomarker is measured with aPCR assay.

In one embodiment, the subject is a mammal.

In one embodiment, the effective amount of the at least one of thegold(III) complex represented by formula (I), the gold(III) complexrepresented by formula (II), the pharmaceutically acceptable salt,solvate, prodrug, and a combination thereof is in a range of 1-100mg/kg.

The second aspect of the disclosure relates to a gold(III) complexrepresented by formula (I) or formula (II):

or a pharmaceutically acceptable salt, solvate, or prodrug thereof;

where each R₁ is independently a hydrogen, an optionally substitutedC₁-C₈ alkyl group, or an optionally substituted C₆-C₈ aryl group;

each R₂ is independently a hydrogen, a halogen, a hydroxyl, an amino, anitro, a cyano, an optionally substituted alkyl, an optionallysubstituted cycloalkyl, an optionally substituted heterocyclyl, anoptionally substituted arylalkyl, an optionally substituted heteroaryl,an optionally substituted alkoxyl, an optionally substituted aryl, anoptionally substituted alkenyl, a N-monosubstituted amino group, or aN,N-disubstituted amino group; and

X is at least one pharmaceutically acceptable anion such as chloride,bromide, and iodide;

with the proviso that R₁ and R₂ are not each a hydrogen.

In one embodiment, at least one R₁ is an optionally substituted C₁-C₈alkyl group.

In one embodiment, at least one R₂ is an optionally substituted alkylgroup.

In one embodiment, the gold(III) complex represented by formula (I) hasX as chloride.

In one embodiment, the gold(III) complex of the second aspect furthercomprises at least one counterion which is at least one pharmaceuticallyacceptable anion selected from the group consisting of fluoride,chloride, bromide, iodide, nitrate, sulfate, phosphate,methanesulfonate, ethanesulfonate, p-toluenesulfonate, salicylate,malate, maleate, succinate, tartrate, citrate, acetate, perchlorate,trifluoromethanesulfonate, acetylacetonate, hexafluorophosphate, andhexafluoroacetylacetonate.

In one embodiment, the at least one counterion is chloride.

BRIEF DESCRIPTION OF THE DRAWINGS

A more complete appreciation of the disclosure and many of the attendantadvantages thereof will be readily obtained as the same becomes betterunderstood by reference to the following detailed description whenconsidered in connection with the accompanying drawings, wherein:

FIG. 1 is a solid-state ¹³C spectrum for complex 2 at a spinning rate of4 kHz.

FIG. 2A is a plot of the observed rate constant against GSHconcentration at various temperatures for the reaction of[(DACH)AuCl₂]Cl with GSH in the presence of 20 mM NaCl.

FIG. 2B is a plot of the observed rate constant against GSHconcentration at various temperatures for the reaction of [(DACH)₂Au]Cl₃with GSH in the presence of 20 mM NaCl.

FIG. 3 is a view of the molecular structure of complex 1 with labelledatoms, and the displacement ellipsoids are drawn at the 50% probabilitylevel.

FIG. 4 shows a crystal packing of complex 1 viewed along the a-axis.

FIG. 5 is a view of the molecular structure of complex 2 with labelledatoms, and the displacement ellipsoids are drawn at the 50% probabilitylevel.

FIG. 6 shows a crystal packing of complex 2 viewed along the a-axis withhydrogen bonds represented by dashed lines, and C-bound H atoms havebeen omitted for clarity.

FIG. 7 shows a crystal packing of complex 2 viewed along the c-axis withhydrogen bonds represented by dashed lines, and C-bound H atoms havebeen omitted for clarity.

FIG. 8A is a micrograph of classical Hodgkin lymphoma L-540 cells in acell culture medium without complex 1 and/or complex 2.

FIG. 8B is a micrograph of L-540 cells after a 72-hour treatment with 10μM of complex 1.

FIG. 8C is a micrograph of L-540 cells after a 72-hour treatment with 25μM of complex 1.

FIG. 8D is a micrograph of L-540 cells after a 72-hour treatment with 50μM of complex 1.

FIG. 8E is a micrograph of L-540 cells after a 72-hour treatment with100 μM of complex 1.

FIG. 8F is a micrograph of L-540 cells after a 72-hour treatment with 10μM of complex 2.

FIG. 8G is a micrograph of L-540 cells after a 72-hour treatment with 25μM of complex 2.

FIG. 8H is a micrograph of L-540 cells after a 72-hour treatment with 50μM of complex 2.

FIG. 8I is a micrograph of L-540 cells after a 72-hour treatment with100 μM of complex 2.

FIG. 8J is a histogram showing the percentage of living L-540 cellsevaluated by a trypan blue dye exclusion assay after a 72-hour treatmentwith complex 1.

FIG. 8K is a histogram showing the percentage of living L-540 cellsevaluated by a trypan blue dye exclusion assay after a 72-hour treatmentwith complex 2.

FIG. 9A is a micrograph of ovarian cancer A2780 cells in a cell culturemedium without complex 1 and/or complex 2.

FIG. 9B is a micrograph of A2780 cells after a 72-hour treatment with 5μM of complex 1.

FIG. 9C is a micrograph of A2780 cells after a 72-hour treatment with 5μM of complex 1.

FIG. 9D is a micrograph of A2780 cells after a 72-hour treatment with 10μM of complex 1.

FIG. 9E is a micrograph of A2780 cells after a 72-hour treatment with 25μM of complex 1.

FIG. 9F is a micrograph of A2780 cells after a 72-hour treatment with 5μM of complex 2.

FIG. 9G is a micrograph of A2780 cells after a 72-hour treatment with 5μM of complex 2.

FIG. 9H is a micrograph of A2780 cells after a 72-hour treatment with 10μM of complex 2.

FIG. 9I is a micrograph of A2780 cells after a 72-hour treatment with 25μM of complex 2.

FIG. 9J is a micrograph of cisplatin-resistant ovarian cancer A2780ciscells in a cell culture medium without complex 1 and/or complex 2.

FIG. 9K is a micrograph of A2780cis cells after a 72-hour treatment with10 μM of complex 1.

FIG. 9L is a micrograph of A2780cis cells after a 72-hour treatment with25 μM of complex 1.

FIG. 9M is a micrograph of A2780cis cells after a 72-hour treatment with50 μM of complex 1.

FIG. 9N is a micrograph of A2780cis cells after a 72-hour treatment with100 μM of complex 1.

FIG. 9O is a micrograph of A2780cis cells after a 72-hour treatment with10 μM of complex 2.

FIG. 9P is a micrograph of A2780cis cells after a 72-hour treatment with25 μM of complex 2.

FIG. 9Q is a micrograph of A2780cis cells after a 72-hour treatment with50 μM of complex 2.

FIG. 9R is a micrograph of A2780cis cells after a 72-hour treatment with100 μM of complex 2.

FIG. 9S is a histogram showing the percentage of living A2780 cellsevaluated by a MTT assay after a 72-hour treatment with complex 1 andcomplex 2.

FIG. 9T is a histogram showing the percentage of living A2780cis cellsevaluated by a MTT assay after a 72-hour treatment with complex 1 andcomplex 2.

DETAILED DESCRIPTION OF THE EMBODIMENTS

Embodiments of the present disclosure will now be described more fullyhereinafter with reference to the accompanying drawings, in which some,but not all of the embodiments of the disclosure are shown.

The present disclosure will be better understood with reference to thefollowing definitions.

As used herein, the words “a” and “an” and the like carry the meaning of“one or more”. Within the description of this disclosure, where anumerical limit or range is stated, the endpoints are included unlessstated otherwise. Also, all values and subranges within a numericallimit or range are specifically included as if explicitly written out.

As used herein, “compound” and “complex” are used interchangeably, andare intended to refer to a chemical entity, whether in the solid, liquidor gaseous phase, and whether in a crude mixture or purified andisolated.

The first aspect of the disclosure relates to a method for treatingcancer, comprising administering an effective amount of at least one ofa gold(III) complex represented by formula (I), a gold(III) complexrepresented by formula (II):

a pharmaceutically acceptable salt, solvate, prodrug, and a combinationthereof to a subject, where the cancer may be ovarian cancer, Hodgkinlymphoma, or both, each of R₁ may be independently a hydrogen, anoptionally substituted C₁-C₈ alkyl group, or an optionally substitutedC₆-C₈ aryl group, each of R₂ may be independently a hydrogen, a halogen,a hydroxyl, an amino, a nitro, a cyano, an optionally substituted alkyl,an optionally substituted cycloalkyl, an optionally substitutedheterocyclyl, an optionally substituted arylalkyl, an optionallysubstituted heteroaryl, an optionally substituted alkoxyl, an optionallysubstituted aryl, an optionally substituted alkenyl, a N-monosubstitutedamino group, or a N,N-disubstituted amino group; and X may be at leastone pharmaceutically acceptable anion such as chloride, bromide, oriodide. In one embodiment, the cancer is not prostate cancer, lungcancer, and/or gastric cancer.

The term “alkyl”, as used herein, unless otherwise specified, refers toa straight or branched hydrocarbon fragment. Non-limiting examples ofsuch hydrocarbon fragments include methyl, ethyl, propyl, isopropyl,butyl, isobutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, isohexyl,3-methylpentyl, 2,2-dimethylbutyl, and 2,3-dimethylbutyl. As usedherein, the term “cycloalkyl” refers to a cyclized alkyl group.Exemplary cycloalkyl groups include, but are not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl, andadamantyl. Branched cycloalkyl groups, for example, 1-methylcyclopropyland 2-methycyclopropyl groups, are included in the definition ofcycloalkyl as used in the present disclosure.

The term “alkenyl” refers to a straight, branched, or cyclic hydrocarbonfragment containing at least one C═C double bond. Exemplary alkenylgroups include, without limitation, 1-propenyl, 2-propenyl (or “allyl”),1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl,4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl,1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 5-heptenyl, 6-heptenyl,1-octenyl, 2-octenyl, 3-octenyl, 4-octenyl, 5-octenyl, 6-octenyl,7-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 4-nonenyl, 5-nonenyl,6-nonenyl, 7-nonenyl, 8-nonenyl, 1-decenyl, 2-decenyl, 3-decenyl,4-decenyl, 5-decenyl, 6-decenyl, 7-decenyl, 8-decenyl, and 9-decenyl.

The term “aryl”, as used herein, and unless otherwise specified, refersto phenyl, biphenyl, naphthyl, anthracenyl, and the like. The term“heteroaryl” refers to an aryl group where at least one carbon atom isreplaced with a heteroatom (e.g. nitrogen, oxygen, sulfur) and can beindolyl, furyl, imidazolyl, triazolyl, triazinyl, oxazolyl, isoxazolyl,thiazolyl, isothiazolyl, pyrazolyl, pyrrolyl, pyrazinyl, tetrazolyl,pyridyl (or its N-oxide), thienyl, pyrimidinyl (or its N-oxide),IH-indolyl, isoquinolyl (or its N-oxide), or quinolyl (or its N-oxide),for example.

As used herein, the term “substituted” refers to at least one hydrogenatom that is replaced with a non-hydrogen group, provided that normalvalencies are maintained and that the substitution results in a stablecompound. When a compound or a R group is noted as “optionallysubstituted”, the substituents are selected from the exemplary groupincluding, but not limited to, aroyl (as defined hereinafter); halogen(e.g. chlorine, bromine, fluorine or iodine); alkoxy (i.e. straight orbranched chain alkoxy having 1 to 10 carbon atoms, and includes, forexample, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy,sec-butoxy, tert-butoxy, pentoxy, isopentoxy, hexyloxy, heptyloxy,octyloxy, nonyloxy, and decyloxy); cycloalkyloxy includingcyclopentyloxy, cyclohexyloxy, and cycloheptyloxy; aryloxy includingphenoxy and phenoxy substituted with halogen, alkyl, alkoxy, andhaloalkyl (which refers to straight or branched chain alkyl having 1 to8 carbon atoms which are substituted by at least one halogen, andincludes, for example, chloromethyl, bromomethyl, fluoromethyl,iodomethyl, 2-chloroethyl, 2-bromoethyl, 2-fluoroethyl, 3-chloropropyl,3-bromopropyl, 3-fluoropropyl, 4-chlorobutyl, 4-fluorobutyl,dichloromethyl, dibromomethyl, difluoromethyl, diiodomethyl,2,2-dichloroethyl, 2,2-dibromoethyl, 2,2-difluoroethyl,3,3-dichloropropyl, 3,3-difluoropropyl, 4,4-dichlorobutyl,4,4-difluorobutyl, trichloromethyl, trifluoromethyl,2,2,2-tri-fluoroethyl, 2,3,3-trifluoropropyl, 1,1,2,2-tetrafluoroethyl,2,2,3,3-tetrafluoropropyl); hydrocarbyl; arylalkyl; hydroxy; alkoxy;oxo; alkanoyl; alkanoyloxy; amino; alkylamino; arylamino;arylalkylamino; disubstituted amines (e.g. in which the two aminosubstituents are selected from the exemplary group including, but notlimited to, alkyl, aryl, or arylalkyl); alkanoylamino; thiol; alkylthio;arylthio; arylalkylthio; alkylthiono; arylthiono; aryalkylthiono;alkylsulfonyl; arylsulfonyl; arylalkylsulfonyl; sulfonamido (e.g.—SO₂NH₂); substituted sulfonamide; nitro; cyano; carboxy; carbamyl (e.g.—CONH₂, —CONHalkyl, —CONHaryl, —CONHarylalkyl or cases where there aretwo substituents on one nitrogen from alkyl, aryl, or arylalkyl);alkoxycarbonyl; aryl; heteroarylcarbonyl; heterocyclyl; and mixturesthereof and the like. The substituents may be either unprotected, orprotected as necessary, as known to those skilled in the art, forexample, as taught in Greene, et al., “Protective Groups in OrganicSynthesis”, John Wiley and Sons, Second Edition, 1991, herebyincorporated by reference in its entirety).

The term “heterocyclyl” as used in this disclosure refers to a 3-8,preferably 4-8, more preferably 4-7 membered monocyclic ring or a fused8-12 membered bicyclic ring which may be saturated or partiallyunsaturated, which monocyclic or bicyclic ring contains 1 to 4heteroatoms selected from oxygen, nitrogen, silicon, or sulfur. Examplesof such monocyclic rings include oxaziridinyl, homopiperazinyl,oxiranyl, dioxiranyl, aziridinyl, pyrrolidinyl, azetidinyl,pyrazolidinyl, oxazolidinyl, piperidinyl, piperazinyl, morpholinyl,thiomorpholinyl, thiazolidinyl, hydantoinyl, valerolactamyl, oxiranyl,oxetanyl, dioxolanyl, dioxanyl, oxathiolanyl, oxathianyl, dithianyl,dihydrofuranyl, tetrahydrofuranyl, dihydropyranyl, tetrahydropyranyl,tetrahydropyridyl, tetrahydropyrimidinyl, tetrahydrothiophenyl,tetrahydrothiopyranyl, diazepanyl, and azepanyl. Examples of suchbicyclic rings include indolinyl, isoindolinyl, benzopyranyl,quinuclidinyl, 2,3,4,5-tetrahydro-1,3,benzazepine,4-(benzo-1,3,dioxol-5-methyl)piperazine, and tetrahydroisoquinolinyl.Further, “substituted heterocyclyl” may refer to a heterocyclyl ringwhich has additional (e.g. one or more) oxygen atoms bonded to the ringatoms of parent heterocylcyl ring. An example of a heterocyclylsubstituted with one or more oxygen atoms is1,1-dioxido-1,3-thiazolidinyl.

The term “alkylthio” as used in this disclosure refers to a divalentsulfur with alkyl occupying one of the valencies and includes the groupsmethylthio, ethylthio, propylthio, butylthio, pentylthio, hexylthio, andoctylthio.

The term “alkanoyl” as used in this disclosure refers to an alkyl grouphaving 2 to 18 carbon atoms that is bound with a double bond to anoxygen atom. Examples of alkanoyl include, acetyl, propionyl, butyryl,isobutyryl, pivaloyl, valeryl, hexanoyl, octanoyl, lauroyl, andstearoyl.

Examples of aroyl are benzoyl and naphthoyl, and “substituted aroyl” mayrefer to benzoyl or naphthoyl substituted by at least one substituentincluding those selected from halogen, amino, nitro, hydroxy, alkyl,alkoxy and haloalkyl on the benzene or naphthalene ring.

The term “arylalkyl” as used in this disclosure refers to a straight orbranched chain alkyl moiety having 1 to 8 carbon atoms that issubstituted by an aryl group or a substituted aryl group having 6 to 12carbon atoms, and includes benzyl, 2-phenethyl, 2-methylbenzyl,3-methylbenzyl, 4-methylbenzyl, 2,4-dimethylbenzyl,2-(4-ethylphenyl)ethyl, 3-(3-propylphenyl)propyl.

The term “heteroarylcarbonyl” as used in this disclosure refers to aheteroaryl moiety with 5 to 10 membered mono- or fused-heteroaromaticring having at least one heteroatom selected from nitrogen, oxygen andsulfur as mentioned above, and includes, for example, furoyl,nicotinoyl, isonicotinoyl, pyrazolylcarbonyl, imidazolylcarbonyl,pyrimidinylcarbonyl, and benzimidazolyl-carbonyl. Further, “substitutedheteroarylcarbonyl” may refer to the above mentioned heteroarylcarbonylwhich is substituted by at least one substituent selected from halogen,amino, vitro, hydroxy, alkoxy and haloalkyl on the heteroaryl nucleus,and includes, for example, 2-oxo-1,3-dioxolan-4-ylmethyl,2-oxo-1,3-dioxan-5-yl.

Vinyl refers to an unsaturated substituent having at least oneunsaturated double bond and having the formula CH₂═CH—. Accordingly,said “substituted vinyl” may refer to the above vinyl substituent havingat least one of the protons on the terminal carbon atom replaced withalkyl, cycloalkyl, cycloalkylalkyl, aryl, substituted aryl, heteroarylor substituted heteroaryl.

The term “hydrocarbyl” as used herein refers to a univalent hydrocarbongroup containing up to about 24 carbon atoms (i.e. a group containingonly carbon and hydrogen atoms) and that is devoid of olefinic andacetylenic unsaturation, and includes alkyl, cycloalkyl,alkyl-substituted cycloalkyl, cycloalkyl-substituted cycloalkyl,cycloalkylalkyl, aryl, alkyl-substituted aryl, cycloalkyl-substitutedaryl, arylalkyl, alkyl-substituted aralkyl, and cycloalkyl-substitutedaralkyl. Further, functionally-substituted hydrocarbyl groups may referto a hydrocarbyl group that is substituted by one or more functionalgroups selected from halogen atoms, amino, nitro, hydroxy,hydrocarbyloxy (including alkoxy, cycloalkyloxy, and aryloxy),hydrocarbylthio (including alkylthio, cycloalkylthio, and arylthio),heteroaryl, substituted heteroaryl, alkanoyl, aroyl, substituted aroyl,heteroarylcarbonyl, and substituted heteroarylcarbonyl.

In some embodiments, R₁ is hydrogen. In one embodiment, R₂ is hydrogen.In another embodiment, X is chloride.

In the gold(III) complex of formula (I), the gold(III) atom may becovalently coordinated to the two vicinal nitrogen atoms on the ligandwith a diaminocyclohexane skeletal structure and the two X ligands. Inthe gold(III) complex of formula (II), the gold(III) atom may becovalently coordinated to the two vicinal nitrogen atoms on each ligand.

In some embodiments, the gold(III) complex represented by formula (I) is

In some embodiments, the gold(III) complex represented by formula (II)is

The generally accepted convention for representing stereochemicalcompounds, which is also adhered to herein, is the following:

a compound represented without stereo bonds, e.g. the cyclohexylfragment, is racemic or the configuration of the stereogenic center isnot defined;

a compound described with one of the descriptors “(±)”, “rel”, or “rac”,is racemic;

a compound represented with solid bars refers to a non-racemic compoundand the stereochemistry of the stereogenic centers are relative; and

a compound represented with solid and broken wedges but without thedescriptors “(±)”, “rel”, or “rac” refers to a non-racemic or anenantio-enriched compound, where its stereochemistry is absolute.

In most embodiments, the gold(III) complex is administered as apharmaceutically acceptable salt having at least one counterion. Theterm “pharmaceutically acceptable salt” refers the gold(III) complex offormula (I) or the gold(III) complex of formula(II) with a counterion.As used herein, the term “counterion” refers to an anion, preferably apharmaceutically acceptable anion that is associated with the gold(III)complex of formula (I) or the gold(III) complex of formula (II).Non-limiting examples of pharmaceutically acceptable counterions includehalides, such as fluoride, chloride, bromide, iodide, nitrate, sulfate,phosphate, amide, methanesulfonate, ethanesulfonate, p-toluenesulfonate,salicylate, malate, maleate, succinate, tartrate, citrate, acetate,perchlorate, trifluoromethanesulfonate (triflate), acetylacetonate,hexafluorophosphate, and hexafluoroacetylacetonate. In some embodiments,the counterion is a halide, preferably chloride.

The phrase “pharmaceutically acceptable” as used herein refers tocounterions, compounds, materials, compositions, and/or dosage formswhich are, within the scope of sound medical judgment, suitable for usein contact with the tissues of human beings and animals withoutexcessive toxicity, irritation, allergic response, or other problem orcomplication, commensurate with a reasonable benefit/risk ratio.Therefore, the composition refers to the combination of an activeingredient with a carrier, inert or active, making the compositionespecially suitable for diagnostic or therapeutic use in vivo or exvivo.

As used herein, “derivative” refers to a chemically or biologicallymodified version of a chemical compound that is structurally similar toa parent compound and (actually or theoretically) derivable from thatparent compound. A “derivative” differs from an “analog” in that aparent compound may be the starting material to generate a “derivative”,whereas the parent compound may not necessarily be used as the startingmaterial to generate an “analog”. A derivative may or may not havedifferent chemical or physical properties of the parent compound. Forexample, the derivative may be more hydrophilic or it may have alteredreactivity compared to the parent compound. Derivatization (i.e.modification) may involve substitution of one or more moieties withinthe molecule (e.g. a change in functional group). The term “derivative”also includes conjugates, and prodrugs of a parent compound (i.e.chemically modified derivatives which can be converted into the originalcompound under physiological conditions).

As used herein, the term “analog” refers to a chemical compound that isstructurally similar to a parent compound, but differs slightly incomposition (e.g. at least one atom or functional group is different,added, or removed). The analog may or may not have different chemical orphysical properties than the original compound and may or may not haveimproved biological and/or chemical activity. For example, the analogmay be more hydrophilic or it may have altered reactivity compared tothe parent compound. The analog may mimic the chemical and/or biologicalactivity of the parent compound (i.e. it may have similar or identicalactivity), or, in some cases, may have increased or decreased activity.The analog may be a naturally or non-naturally occurring variant of theoriginal compound. Other types of analogs include isomers (enantiomers,diastereomers, and the like) and other types of chiral variants of acompound, as well as structural isomers.

The term “solvate” means a physical association of the gold(III) complexof formula (I) or the gold(III) complex of formula (II) with one or moresolvent molecules, whether organic or inorganic. This physicalassociation includes hydrogen bonding. In certain instances the solvatewill be capable of isolation, for example when one or more solventmolecules are incorporated in the crystal lattice of the crystallinesolid. The solvent molecules in the solvate may be present in a regulararrangement and/or a non-ordered arrangement. The solvate may compriseeither a stoichiometric or non-stoichiometric amount of the solventmolecules. Solvate encompasses both solution-phase and isolablesolvates. Exemplary solvates include, but are not limited to, hydrates,ethanolates, methanolates, and isopropanolates. Methods of solvation aregenerally known in the art.

The present disclosure is further intended to include all isotopes ofatoms occurring in the present compounds. Isotopes include those atomshaving the same atomic number but different mass numbers. By way ofgeneral example, and without limitation, isotopes of hydrogen includedeuterium and tritium. Isotopes of carbon include ¹³C and ¹⁴C.Isotopically labeled compounds of the disclosure can generally beprepared by conventional techniques known to those skilled in the art orby processes and methods analogous to those described herein, using anappropriate isotopically labeled reagent in place of the non-labeledreagent otherwise employed.

As used herein, the terms “treat”, “treatment”, and “treating” in thecontext of the administration of a therapy to a subject in need thereofrefer to the reduction or inhibition of the progression and/or durationof a disease (e.g. cancer), the reduction or amelioration of theseverity of the disease, and/or the amelioration of one or more symptomsthereof resulting from the administration of one or more therapies.“Treating” or “treatment” of the disease includes preventing the diseasefrom occurring in a subject that may be predisposed to the disease butdoes not yet experience or exhibit symptoms of the disease (prophylactictreatment), inhibiting the disease (slowing or arresting itsdevelopment), ameliorating the disease, providing relief from thesymptoms or side-effects of the disease (including palliativetreatment), and relieving the disease (causing regression of thedisease). With regard to the disease, these terms simply mean that oneor more of the symptoms of the disease will be reduced. Such terms mayrefer to one, two, three, or more results following the administrationof one, two, three, or more therapies: (1) a stabilization, reduction(e.g. by more than 10%, 20%, 30%, 40%, 50%, preferably by more than 60%of the population of cancer cells and/or tumor size beforeadministration), or elimination of the cancer cells, (2) inhibitingcancerous cell division and/or cancerous cell proliferation, (3)relieving to some extent (or, preferably, eliminating) one or moresymptoms associated with a pathology related to or caused in part byunregulated or aberrant cellular division, (4) an increase indisease-free, relapse-free, progression-free, and/or overall survival,duration, or rate, (5) a decrease in hospitalization rate, (6) adecrease in hospitalization length, (7) eradication, removal, or controlof primary, regional and/or metastatic cancer, (8) a stabilization orreduction (e.g. by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%,preferably at least 80% relative to the initial growth rate) in thegrowth of a tumor or neoplasm, (9) an impairment in the formation of atumor, (10) a reduction in mortality, (11) an increase in the responserate, the durability of response, or number of patients who respond orare in remission, (12) the size of the tumor is maintained and does notincrease or increases by less than 10%, preferably less than 5%,preferably less than 4%, preferably less than 2%, (13) a decrease in theneed for surgery (e.g. colectomy, mastectomy), and (14) preventing orreducing (e.g. by more than 10%, more than 30%, preferably by more than60% of the population of metastasized cancer cells beforeadministration) the metastasis of cancer cells.

The terms “patient”, “subject”, and “individual” are usedinterchangeably. As used herein, they refer to individuals sufferingfrom a disease and encompass mammals. None of the terms require that theindividual be under the care and/or supervision of a medicalprofessional. Mammals are any member of the mammalian class, includingbut are not limited to humans, non-human primates, such as chimpanzees,and other apes and monkey species, farm animals, such as cattle, horses,sheep, goats, swine, domestic animals, such as rabbits, dogs, and cats,laboratory animals including rodents, such as rats, mice and guineapigs, and the like. In preferred embodiments, the subject is a human.

A subject in need of treatment includes a subject already with thedisease, a subject which does not yet experience or exhibit symptoms ofthe disease, and a subject predisposed to the disease. In preferredembodiments, the subject is a person who is predisposed to cancer, e.g.a person with a family history of cancer. Women who have (i) certaininherited genes (e.g. mutated BRCA1 and/or mutated BRCA2), (ii) beentaking estrogen alone (without progesterone) after menopause for manyyears (at least 5, at least 7, or at least 10), and/or (iii) been takingfertility drug clomiphene citrate, are at a higher risk of contractingovarian cancer. People who have had infectious mononucleosis (aninfection caused by the Epstein-Barr virus (EBV)), and/or are infectedwith HIV (human immunodeficiency virus), are at a higher risk ofcontracting Hodgkin lymphoma.

In another embodiment, the subject refers to a cancer patient who hasbeen previously administered/treated with cisplatin and have cisplatinresistance (for example in the form of high ERCC1 mRNA levels,overexpression of HER-2/neu, activation of the PI3-K/Akt pathway, lossof p53 function, and/or overexpression of antiapoptotic bcl-2).

The methods for treating cancer and other proliferative disordersdescribed herein inhibit, remove, eradicate, reduce, regress, diminish,arrest or stabilize a cancerous tumor, including at least one of thetumor growth, tumor cell viability, tumor cell division andproliferation, tumor metabolism, blood flow to the tumor and metastasisof the tumor. In some embodiments, the size of a tumor, whether byvolume, weight or diameter, is reduced after the treatment by at least5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%,95%, 99%, or 100%, relative to the tumor size before treatment. In otherembodiments, the size of a tumor after treatment does not reduce but ismaintained the same as the tumor size before treatment. Methods ofassessing tumor size include but are not limited to CT scan, MRI,DCE-MRI and PET scan.

As used herein, the terms “therapies” and “therapy” can refer to anymethod, composition, and/or active ingredient that can be used in thetreatment and/or management of the disease or one or more symptomsthereof. In some embodiments, the method for treating the diseaseinvolves the administration of a unit dosage or a therapeuticallyeffective amount of the active ingredient to a subject in need thereof.

The terms “effective amount”, “therapeutically effective amount”, or“pharmaceutically effective amount” refer to that amount of the activeingredient being administered which will relieve to some extent one ormore of the symptoms of the disease being treated. The result can bereduction and/or alleviation of the signs, symptoms, or causes of adisease, or any other desired alteration of a biological system. Forexample, an “effective amount” for therapeutic uses is the amount of thegold(III) complex of formula (I), the gold(III) complex of formula (II),the salt thereof, the solvate thereof, the prodrug thereof, or acombination thereof as disclosed herein required to provide a clinicallysignificant decrease in a disease. An appropriate “effective amount” maydiffer from one individual to another. An appropriate “effective amount”in any individual case may be determined using techniques, such as adose escalation study.

The dosage and treatment duration are dependent on factors, such asbioavailability of a drug, administration mode, toxicity of a drug,gender, age, lifestyle, body weight, the use of other drugs and dietarysupplements, the disease stage, tolerance and resistance of the body tothe administered drug, etc., and then determined and adjustedaccordingly. In at least one embodiment, the at least one of thegold(III) complex of formula (I), the gold(III) complex of formula (II),the salt thereof, the solvate thereof, the prodrug thereof, and thecombination thereof is administered in an effective amount in a range of1-100 mg/kg based on the weight of the subject, preferably 10-80 mg/kg,more preferably 20-50 mg/kg.

In most embodiments, the gold(III) complex of formula (I), the gold(III)complex of formula(II), the salt thereof, the solvate thereof, theprodrug thereof, or the combination thereof is formulated in acomposition. As used herein, a “composition” refers to a mixture of theactive ingredient with other chemical components, such aspharmaceutically acceptable carriers and excipients. One purpose of acomposition is to facilitate administration of the gold(III) complex offormula (I), the gold(III) complex of formula(II), the salt thereof, thesolvate thereof, the prodrug thereof, or a combination thereof to asubject. Depending on the intended mode of administration (oral,parenteral, or topical), the composition can be in the form of solid,semi-solid or liquid dosage forms, such as tablets, suppositories,pills, capsules, powders, liquids, or suspensions, preferably in unitdosage form suitable for single administration of a precise dosage.

The term “active ingredient”, as used herein, refers to an ingredient inthe composition that is biologically active, for example, the gold(III)complex of formula (I), the gold(III) complex of formula (II), a saltthereof, a prodrug thereof, and a solvate thereof.

In most embodiments, the composition comprises at least 0.5 wt %, 5 wt%, 10 wt %, 15 wt %, 20 wt %, 25 wt %, 30 wt %, 35 wt %, 40 wt %, 45 wt%, 50 wt %, 55 wt %, 60 wt %, 65 wt %, 70 wt %, 75 wt %, 80 wt %, 85 wt%, 90 wt %, 95 wt %, 99 wt %, or 99.9 wt %, of the gold(III) complex offormula (I), the gold(III) complex of formula (II), the pharmaceuticallyacceptable salt thereof, the pharmaceutically acceptable solvatethereof, or a combination thereof. The composition may comprise 0.01-50μM, 0.01-30 μM, preferably 0.01-10 μM of the gold(III) complex offormula (I) or the gold(III) complex of formula (II) relative to thetotal composition. In some embodiments, the composition comprises up to0.1 wt %, 1 wt %, 5 wt %, or 10 wt % of the pharmaceutically acceptablesalt of either the gold(III) complex of formula (I) or the gold(III)complex of formula (II). In some embodiments, the composition comprisesup to 0.1 wt %, 1 wt %, 5 wt %, or 10 wt % of the pharmaceuticallyacceptable solvate thereof of either the gold(III) complex of formula(I) or the gold(III) complex of formula (II). Preferably, thecomposition may further comprise pharmaceutically acceptable binders,such as sucrose, lactose, xylitol, and pharmaceutically acceptableexcipients such as calcium carbonate, calcium phosphate, and dimethylsulfoxide (DMSO).

The neoplastic activity of the tumor or cancer cells may be localized orinitiated in one or more of the following: blood, brain, bladder, lung,cervix, ovary, colon, rectum, pancreas, skin, prostate gland, stomach,intestine, breast, liver, spleen, kidney, head, neck, testicle, bone(including bone marrow), thyroid gland, and central nervous system.Preferably, the composition may be used to treat ovarian cancer andHodgkin lymphoma such as classical Hodgkin lymphoma and nodularlymphocyte-predominant Hodgkin lymphoma. In some embodiments, thecomposition is used to treat cisplatin-resistant ovarian cancer and/orclassical Hodgkin lymphoma. Therefore, in one embodiment, the subjecthas ovarian cancer and is currently undergoing, or has completed acisplatin-based chemotherapy regimen.

In treating certain cancers, the best approach is a combination ofsurgery, radiotherapy, and/or chemotherapy. Therefore, in at least oneembodiment, the composition is employed with radiotherapy. In anotherembodiment, the composition is employed with surgery. The radiotherapyand/or surgery may be before or after the composition is administered.

In one embodiment, the composition is used for treating cancer andfurther comprises a second active ingredient, such as a chemotherapeuticagent, for the treatment or prevention of neoplasm, of tumor or cancercell division, growth, proliferation and/or metastasis in the subject;induction of death or apoptosis of tumor and/or cancer cells; and/or anyother form of proliferative disorder. In some embodiments, the firstactive ingredient is the gold(III) complex of formula (I) and the secondactive ingredient is the gold(III) complex of formula (II), or viceversa.

Exemplary chemotherapeutic agents include, without limitation,aflibercept, asparaginase, bleomycin, busulfan, carmustine,chlorambucil, cladribine, cyclophosphamide, cytarabine, dacarbazine,daunorubicin, doxorubicin, etoposide, fludarabine, gemcitabine,hydroxyurea, idarubicin, ifosfamide, irinotecan, lomustine,mechlorethamine, melphalan, mercaptopurine, methotrexate, mitomycin,mitoxantrone, pentostatin, procarbazine, topotecan, vinblastine,vincristine, retinoic acid, oxaliplatin, carboplatin, 5-fluorouracil,teniposide, amasacrine, docetaxel, paclitaxel, vinorelbine, bortezomib,clofarabine, capecitabine, actinomycin D, epirubicin, vindesine,methotrexate, 6-thioguanine, tipifarnib, imatinib, erlotinib, sorafenib,sunitinib, dasatinib, nilotinib, lapatinib, gefitinib, temsirolimus,everolimus, rapamycin, bosutinib, pzopanib, axitinib, neratinib,vatalanib, pazopanib, midostaurin, enzastaurin, trastuzumab, cetuximab,panitumumab, rituximab, bevacizumab, mapatumumab, conatumumab, andlexatumumab. The composition may comprise 0.1-50 wt % of the secondactive ingredient, preferably 10-40 wt %, more preferably 10-20 wt %,relative to the weight of the first active ingredient.

As used herein, a “pharmaceutically acceptable carrier” refers to acarrier or diluent that does not cause significant irritation to anorganism, does not abrogate the biological activity and properties ofthe administered active ingredient, and/or does not interact in adeleterious manner with the other components of the composition in whichit is contained. The term “carrier” encompasses any excipient, binder,diluent, filler, salt, buffer, solubilizer, lipid, stabilizer, or othermaterial well known in the art for use in pharmaceutical formulations.The choice of a carrier for use in a composition will depend upon theintended route of administration for the composition. The preparation ofpharmaceutically acceptable carriers and formulations containing thesematerials is described in, e.g. Remington's Pharmaceutical Sciences,21st Edition, ed. University of the Sciences in Philadelphia,Lippincott, Williams & Wilkins, Philadelphia Pa., 2005, which isincorporated herein by reference in its entirety). Examples ofphysiologically acceptable carriers include buffers such as phosphatebuffers, citrate buffer, and buffers with other organic acids;antioxidants including ascorbic acid; low molecular weight (less thanabout 10 residues) polypeptides; proteins, such as serum albumin,gelatin, or immunoglobulins; hydrophilic polymers such aspolyvinylpyrrolidone; amino acids such as glycine, glutamine,asparagine, arginine or lysine; monosaccharides, disaccharides, andother carbohydrates including glucose, mannose, or dextrins; chelatingagents such as EDTA; sugar alcohols such as mannitol or sorbitol;salt-forming counterions such as sodium; and/or nonionic surfactantssuch as TWEEN® (ICI, Inc.; Bridgewater, N.J.), polyethylene glycol(PEG), and PLURONICS™ (BASF; Florham Park, N.J.). An “excipient” refersto an inert substance added to a composition to further facilitateadministration of a compound. Examples, without limitation, ofexcipients include calcium carbonate, calcium phosphate, various sugarsand types of starch, cellulose derivatives, gelatin, vegetable oils, andpolyethylene glycols.

The composition thereof may be administered in a single dose or multipleindividual divided doses. In some embodiments, the composition isadministered at various dosages (e.g. a first dose with an effectiveamount of 50 mg/kg and a second dose with an effective amount of 10mg/kg). In some embodiments, the interval of time between theadministration of the composition and the administration of one or moreadditional therapies may be about 1-5 minutes, 1-30 minutes, 30 minutesto 60 minutes, 1 hour, 1-2 hours, 2-6 hours, 2-12 hours, 12-24 hours,1-2 days, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 1 week, 2weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10weeks, 15 weeks, 20 weeks, 26 weeks, 52 weeks, 11-15 weeks, 15-20 weeks,20-30 weeks, 30-40 weeks, 40-50 weeks, 1 month, 2 months, 3 months, 4months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11months, 12 months, 1 year, 2 years, or any period of time in between.Preferably, the composition is administered once daily for at least 2days, 5 days, 6 days, or 7 days. In certain embodiments, the compositionand one or more additional therapies are administered less than 1 day, 1week, 2 weeks, 3 weeks, 4 weeks, one month, 2 months, 3 months, 6months, 1 year, 2 years, or 5 years apart.

The terms “administer”, “administering”, “administration”, and the like,as used herein, refer to the methods that may be used to enable deliveryof the active ingredient and/or the composition to the desired site ofbiological action. Routes or modes of administration are as set forthherein. These methods include, but are not limited to, oral routes,intraduodenal routes, parenteral injection (including intravenous,subcutaneous, intraperitoneal, intramuscular, intravascular, orinfusion), topical and rectal administration. Those of ordinary skill inthe art are familiar with administration techniques that can be employedwith the compounds and methods described herein. In preferredembodiments, the active ingredient and/or the composition describedherein are administered orally.

In other embodiments, the composition has various release rates (e.g.controlled release or immediate release). Immediate release refers tothe release of an active ingredient substantially immediately uponadministration. In another embodiment, immediate release occurs whenthere is dissolution of an active ingredient within 1-20 minutes afteradministration. Dissolution can be of all or less than all (e.g. about70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%,about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about99%, about 99.5%, 99.9%, or 99.99%) of the active ingredient. In anotherembodiment, immediate release results in complete or less than completedissolution within about 1 hour following administration. Dissolutioncan be in a subject's stomach and/or intestine. In one embodiment,immediate release results in dissolution of an active ingredient within1-20 minutes after entering the stomach. For example, dissolution of100% of an active ingredient can occur in the prescribed time. Inanother embodiment, immediate release results in complete or less thancomplete dissolution within about 1 hour following rectaladministration. In some embodiments, immediate release is throughinhalation, such that dissolution occurs in a subject's lungs.

Controlled-release, or sustained-release, refers to the release of anactive ingredient from a composition or dosage form in which the activeingredient is released over an extended period of time. In oneembodiment, controlled-release results in dissolution of an activeingredient within 20-180 minutes after entering the stomach. In anotherembodiment, controlled-release occurs when there is dissolution of anactive ingredient within 20-180 minutes after being swallowed. Inanother embodiment, controlled-release occurs when there is dissolutionof an active ingredient within 20-180 minutes after entering theintestine. In another embodiment, controlled-release results insubstantially complete dissolution after at least 1 hour followingadministration. In another embodiment, controlled-release results insubstantially complete dissolution after at least 1 hour following oraladministration. In another embodiment, controlled-release results insubstantially complete dissolution after at least 1 hour followingrectal administration. In one embodiment, the composition is not acontrolled-release composition.

Solid dosage forms for oral administration can include capsules,tablets, pills, powders, and granules. In such solid dosage forms, theactive ingredient is ordinarily combined with one or more adjuvantsappropriate to the indicated route of administration. If administeredper os, the active ingredient can be admixed with lactose, sucrose,starch powder, cellulose esters of alkanoic acids, cellulose alkylesters, talc, stearic acid, magnesium stearate, magnesium oxide, sodiumand calcium salts of phosphoric and sulfuric acids, gelatin, acacia gum,sodium alginate, polyvinylpyrrolidone, and/or polyvinyl alcohol, andthen tableted or encapsulated for convenient administration. Suchcapsules or tablets can contain a controlled-release formulation as canbe provided in a dispersion of active compound in hydroxypropylmethylcellulose. In the case of capsules, tablets, and pills, the dosage formscan also comprise buffering ingredients such as sodium citrate,magnesium or calcium carbonate or bicarbonate. Tablets and pills canadditionally be prepared with enteric coatings.

Liquid dosage forms for oral administration can include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups, and elixirscontaining inert diluents commonly used in the art, such as water. Suchcompositions can also comprise adjuvants, such as wetting ingredients,emulsifying and suspending ingredients, and sweetening, flavoring, andperfuming ingredients.

For therapeutic purposes, formulations for parenteral administration canbe in the form of aqueous or non-aqueous isotonic sterile injectionsolutions or suspensions. The term “parenteral”, as used herein,includes intravenous, intravesical, intraperitoneal, subcutaneous,intramuscular, intralesional, intracranial, intrapulmonal, intracardial,intrasternal, and sublingual injections, or infusion techniques. Thesesolutions and suspensions can be prepared from sterile powders orgranules having one or more of the carriers or diluents mentioned foruse in the formulations for oral administration. The active ingredientcan be dissolved in water, polyethylene glycol, propylene glycol,ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzylalcohol, sodium chloride, and/or various buffers. Other adjuvants andmodes of administration are well and widely known in the pharmaceuticalart.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions can be formulated according to the known artusing suitable dispersing or wetting ingredients and suspendingingredients. The sterile injectable preparation can also be a sterileinjectable solution or suspension in a non-toxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that can be employed are water,Ringer's solution, and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil can be employedincluding synthetic mono- or diglycerides. In addition, fatty acids,such as oleic acid, find use in the preparation of injectables. Dimethylacetamide, surfactants including ionic and non-ionic detergents,polyethylene glycols can be used. Mixtures of solvents and wettingingredients such as those discussed above are also useful.

Suppositories for rectal administration can be prepared by mixing theactive ingredient with a suitable non-irritating excipient, such ascocoa butter, synthetic mono-, di-, or triglycerides, fatty acids, andpolyethylene glycols that are solid at ordinary temperatures but liquidat the rectal temperature and will therefore melt in the rectum andrelease the drug.

Topical administration can also involve the use of transdermaladministration such as transdermal patches or iontophoresis devices.Formulation of drugs is discussed in, for example, Hoover, John E.,Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa.,1975. Another example of includes Liberman, H. A. and Lachman, L., Eds.,Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980, whichis incorporated herein by reference in its entirety).

In one embodiment, the IC₅₀ of the gold(III) complexes of formula (I) orformula (II) is in a range of 0.01-100 μM, 1-100 μM, 10-90 μM, 20-80 μM,30-80 μM, 40-80 μM, 50-80 μM, or 50-75 μM. As used herein, the term“IC₅₀” refers to a concentration of the gold(III) complex of formula(I), the gold(II) complex of formula (II), the salt thereof, the prodrugthereof, or the solvate thereof, which causes the death of 50% of cancercells in 72 hours (3 days).

The IC₅₀ can be determined by standard cell viability assays, such as,without limitation, ATP test, Calcein AM assay, clonogenic assay,ethidium homodimer assay, Evans blue assay, Fluorescein diacetatehydrolysis/propidium iodide staining assay, flow cytometry assay,formazan-based assays (MTT, XTT), green fluorescent protein assay,lactate dehydrogenase assay, methyl violet assay, propidium iodideassay, Resazurin assay, Trypan Blue assay and TUNEL assay. Preferably, aMTT assay and/or a Trypan Blue assay is used.

In at least one embodiment, the human cancer cells are derived fromcommercial cell lines, including but are not limited to HeLa cervicalcancer cells, A549 lung cancer cells, HCT15 colon cancer cells, HCT8 orHRT8 colon cancer cells, HCT116 colon cancer cells, DLD1 colon cancercells, MCF7 breast cancer cells, MDA-MB231 breast cancer cells, A2780ovarian cancer cells, HePG2 liver cancer cells, L540 Hodgkin lymphomacells, and DU145 prostatic cancer cells. In some embodiments,cisplatin-resistant cancer cells are used. These cells may be culturedwith low doses of cisplatin in order to build resistance to cisplatinwhile maintaining cell viability. Examples of cisplatin-resistant cancercells include, but are not limited to, A2780cis cisplatin-resistantovarian cancer cells and SGC7901cis cisplatin-resistant gastrointestinalcancer cells. In other embodiments, the human cancer cells are cancercells of a human patient who has been diagnosed with, is suspected ofhaving, or is susceptible to or at risk of having at least one form ofcancer, preferably ovarian cancer.

In most embodiments, the method further comprises measuring aconcentration of a biomarker and/or detecting a mutation in a biomarkerbefore and/or after the composition is administered. As used herein, theterm “biomarker” refers to a characteristic that is objectively measuredand evaluated as an indicator of normal biological processes, pathogenicprocesses or pharmacological responses to a therapeutic intervention.Exemplary cancer biomarkers for ovarian cancer and/or Hodgkin lymphomainclude, without limitation, BRCA1, BRCA2, CCL17, CD163, CD30, NF-κB,Gal-1, CA125, HE4, mesothelin, transthyretin, ApoA1, VCAM, IL-6, IL-8,B7-H4, serum amyloid A, transferrin, osteopontin, kallikreins, OVX1,VEGF, AGR-2, inhibin, M-CSF, uPAR, EGF receptor, lysophosphatidyl acid,beta2-microglobulin, miRNA, and Epstein-Barr virus (EBV) DNA.Specifically, potentially predictive cancer biomarkers include, withoutlimitation, mutations in genes BRCA1 and BRCA2 for ovarian cancer.

Cancer biomarkers may be useful in determining the aggressiveness of anidentified cancer as well as its likelihood of responding to thetreatment. Examples of such prognostic biomarkers include, withoutlimitation, CA125, beta2-microglobulin, and EBV DNA.

The mutation in the biomarker may be detected with a polymerase chainreaction (PCR) assay, DNA microarray, multiplex ligation-dependent probeamplification (MLPA), single strand conformational polymorphism (SSCP),denaturing gradient gel electrophoresis (DGGE), heteroduplex analysis,and restriction fragment length polymorphism (RFLP). The procedures todetect the mutation are well-known to those of ordinary skill in theart.

The concentration of the biomarker may be measured with an assay, forexample an antibody-based method (e.g. an ELISA).

As used herein, the term “antibody-based method” refers to any methodwith the use of an antibody including, but not limited to, enzyme-linkedimmunosorbent assay (ELISA), Western blotting, immunoprecipitation (IP),enzyme linked immunospot (ELISPOT), immunostaining,immunohistochemistry, immunocytochemistry, affinity chromatography, andthe like.

Preferably, an ELISA is used. The term “ELISA” refers to a method ofdetecting the presence and concentration of a biomarker in a sample.There are several variants of ELISA, including, but not limited to,sandwich ELISA, competitive ELISA, indirect ELISA, ELISA reverse, andthe like. The ELISA assay may be a singleplex assay or a multiplexassay, which refers to a type of assay that simultaneously measuresmultiple analytes in a single run/cycle of the assay. Preferably, asandwich ELISA is used.

The protocol for measuring the concentration of the biomarker and/ordetecting the mutation in the biomarker is known to those of ordinaryskill, for example by performing the steps outlined in the commerciallyavailable assay kit sold by Sigma-Aldrich, Thermo Fisher Scientific, R &D Systems, ZeptoMetrix Inc., Cayman Inc., Abcam, Trevigen, DojindoMolecular Technologies, Biovision, and Enzo Life Sciences.

The term “sample” includes any biological sample taken from the subjectincluding a cell, tissue sample, or body fluid. For example, a samplemay include a skin sample, a cheek cell sample, saliva, or blood cells.A sample can include, without limitation, a single cell, multiple cells,fragments of cells, an aliquot of a body fluid, whole blood, platelets,serum, plasma, red blood cells, white blood cells, endothelial cells,tissue biopsies, synovial fluid, and lymphatic fluid. In someembodiments, the sample is taken from a tumor.

In some embodiments, the concentration of the biomarker is measuredbefore and after the administration. When the concentration of thebiomarker is maintained, the method may further comprise increasing theeffective amount of at least one of the gold(III) complex of formula(I), the gold(III) complex of formula (II), the salt thereof, thesolvate thereof, the prodrug thereof, and the combination thereof by atleast 5%, at least 10%, or at least 30%, up to 50%, up to 60%, or up to80% of an initial effective amount that is in a range of 1-100 mg/kgbased on the weight of the subject. The increased effective amount maybe in a range of 1.05-180 mg/kg, preferably 15-140 mg/kg, morepreferably 25-90 mg/kg. The subject may be administered with theincreased dosage for a longer period (e.g. 1 week more, 2 weeks more, or2 months more) than the duration with the initial effective amount.

In some embodiments, the mutation in the biomarker is detected beforeadministrating the composition to identify subjects predisposed to thedisease. For example, women with a BRCA1 germline mutation are at ahigher risk of contracting ovarian cancer.

In some embodiments, the biomarkers are measured/detected after eachadministration. For example, the measurement may be 1-5 minutes, 1-30minutes, 30-60 minutes, 1-2 hours, 2-12 hours, 12-24 hours, 1-2 days,1-15 weeks, 15-20 weeks, 20-30 weeks, 30-40 weeks, 40-50 weeks, 1 year,2 years, or any period of time in between after the administration.

In some embodiments, the administration is stopped once the subject istreated.

The second aspect of the disclosure relates to a gold(III) complexrepresented by formula (I) or formula (II):

where each of R₁ may independently be a hydrogen, an optionallysubstituted C₁-C₈ alkyl group, or an optionally substituted C₆-C₈ arylgroup, each of R₂ may independently be a hydrogen, a halogen, ahydroxyl, an amino, a nitro, a cyano, an optionally substituted alkyl,an optionally substituted cycloalkyl, an optionally substitutedheterocyclyl, an optionally substituted arylalkyl, an optionallysubstituted heteroaryl, an optionally substituted alkoxyl, an optionallysubstituted aryl, an optionally substituted alkenyl, a N-monosubstitutedamino group, or a N,N-disubstituted amino group, and X may be at leastone pharmaceutically acceptable anion such as chloride, bromide, andiodide, with the proviso that R₁ and R₂ are not each a hydrogen.

In some embodiments, at least one R₁ is an optionally substituted C₁-C₈alkyl group. For example, at least one R₁ is a methyl group and theremaining R₁ that are not methyl groups are each a hydrogen atom.Further, there may be one methyl group and a hydrogen atom on eachnitrogen atom of the amino group. In one embodiment, at least one R₂ isan optionally substituted alkyl group. For example, at least one R₂ is amethyl group and the remaining R₂ that are not methyl groups are each ahydrogen atom. Further, there may be one methyl group in eachcyclohexyl. In a preferred embodiment, X is chloride.

In most embodiments, the gold(III) complex is in the form of thepharmaceutical acceptable salt having at least one counterion which isat least one pharmaceutically acceptable anion selected from the groupconsisting of fluoride, chloride, bromide, iodide, nitrate, sulfate,phosphate, methanesulfonate, ethanesulfonate, p-toluenesulfonate,salicylate, malate, maleate, succinate, tartrate, citrate, acetate,perchlorate, trifluoromethanesulfonate, acetylacetonate,hexafluorophosphate, and hexafluoroacetylacetonate. Preferably, thecounterion is chloride.

The gold(III) complex represented by formula (I) or formula (II) may beprepared by mixing a gold(III) precursor with a ligand of the followingformula:

Exemplary gold(III) precursors include, without limitation, sodiumtetrachloroaurate(III), potassium tetrachloroaurate(III), cesiumtetrachloroaurate(III), sodium tetrabromoaurate(III), potassiumtetrabromoaurate(III), caesium tetrabromoaurate(III), and hydratesthereof. The gold(III) precursor and the ligand (for example(1R,2R)-(−)-1,2-diaminocyclohexane) may be dissolved separately in asolvent to form a gold(III) precursor solution and a ligand solution. Aconcentration of the gold(III) precursor solution and/or the ligandsolution may be in a range of 0.005-1 M, 0.01-0.5 M, or 0.01-0.1 M. Insome embodiments, the molar ratio of the gold(III) precursor to theligand is in a range of 1:0.9 to 1:1.1, 1:0.95 to 1:1.05, or 1:0.99 to1:1.01 to prepare the gold(III) complex of formula (I). In someembodiments, the molar ratio of the gold(III) precursor to the ligand isin a range of 1:1.9 to 1:2.1, 1:1.95 to 1:2.05, or 1:1.99 to 1:2.01 toprepare the gold(III) complex of formula (II). The ligand solution maybe added dropwise (or at a rate of 0.05-0.5 ml/min, 0.05-0.3 ml/min, or0.05-0.1 ml/min) to the gold(III) precursor solution to form a reactionmixture. The reaction mixture may be shaken/stirred throughout theduration of the reaction by employing a rotary shaker, a magneticstirrer, or an overhead stirrer. In another embodiment, the reactionmixture is left to stand (i.e. not stirred). In one embodiment, thereaction mixture is preferably mixed in a centrifugal mixer with arotational speed of at least 500 rpm, preferably at least 800 rpm, morepreferably at least 1,000 rpm, even though it can also be mixed with aspatula. In one embodiment, the reaction mixture is sonicated. A clearsolution may form and may be cooled at a temperature in a range of 0-10°C., 0-6° C., or 0-4° C., for 1-10 days, 1-5 days, or 1-3 days. Thesolution may be cooled with an external cooling source such as an icebath with or without salt, a thermostatted thermocirculator, or byrefrigerating the solution. Crystallization of the gold(III) complex offormula (I) or formula (II) may occur and the crystals may be collectedusing methods known to those skilled in the art such as filtration.

As used herein, the term “solvent” includes, but is not limited to,water (e.g. tap water, distilled water, doubly distilled water,deionized water, deionized distilled water), organic solvents, such asethers (e.g. diethyl ether, tetrahydrofuran, 1,4-dioxane,tetrahydropyran, t-butyl methyl ether, cyclopentyl methyl ether,di-iso-propyl ether), glycol ethers (e.g. 1,2-dimethoxyethane, diglyme,triglyme), alcohols (e.g. methanol, ethanol, trifluoroethanol,n-propanol, i-propanol, n-butanol, i-butanol, t-butanol, n-pentanol,i-pentanol, 2-methyl-2-butanol, 2-trifluoromethyl-2-propanol,2,3-dimethyl-2-butanol,3-pentanol, 3-methyl-3-pentanol,2-methyl-3-pentanol, 2-methyl-2-pentanol, 2,3-dimethyl-3-pentanol,3-ethyl-3-pentanol, 2-methyl-2-hexanol, 3-hexanol, cyclopropylmethanol,cyclopropanol, cyclobutanol, cyclopentanol, cyclohexanol), aromaticsolvents (e.g. benzene, o-xylene, m-xylene, p-xylene, and mixtures ofxylenes, toluene, mesitylene, anisole, 1,2-dimethoxybenzene,α,α,α,-trifluoromethylbenzene, fluorobenzene), chlorinated solvents(e.g. chlorobenzene, dichloromethane, 1,2-dichloroethane,1,1-dichloroethane, chloroform), ester solvents (e.g. ethyl acetate,propyl acetate), amide solvents (e.g. dimethylformamide,dimethylacetamide, N-methyl-2-pyrrolidone), urea solvents, ketones (e.g.acetone, butanone), acetonitrile, propionitrile, butyronitrile,benzonitrile, dimethyl sulfoxide, ethylene carbonate, propylenecarbonate, 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone, andmixtures thereof. Preferably, the solvent is absolute ethanol.

Having generally described this disclosure, a further understanding canbe obtained by reference to certain specific examples which are providedherein for purposes of illustration only and are not intended to belimiting unless otherwise specified.

Example 1 Chemicals

Sodium tetrachloroaurate(III) dihydrate,(1R,2R)-(−)-1,2-diaminocyclohexane (DACH), and glutathione (GSH) wereobtained from Sigma-Aldrich, USA. Absolute ethanol was obtained fromMerck, D₂O was purchased from Alfa Aesar, and deionized water with aresistivity of 18.6 MΩ cm⁻¹ for solution preparation was obtaineddirectly from a PURELABs Ultra Laboratory Water Purification System.

Example 2 Synthesis of Complexes 1(R),2(R)—[Au(DACH)AuCl₂]Cl (1) and1(R),2(R)—[Au(DACH)₂Au]Cl₃ (2)

1(R),2(R)-[(DACH)AuCl₂]Cl (1) was synthesized by the direct mixing of200 mg (0.5 mmol) of NaAuCl₄.2H₂O with 57 mg (0.5 mmol) of(1R,2R)-(−)-1,2-Diaminocyclohexane ligand in alcoholic media aspreviously described (S. S. Al-Jaroudi, M. Fettouhi, M. I. M. Wazeer, A.A. Isab, and S. Altuwaijri, Polyhedron, vol. 50, no. 1, pp. 434-442,2013, incorporated herein by reference in its entirety). First, 200 mg(0.5 mmol) NaAuCl₄.2H₂O was dissolved using absolute ethanol, 57 mg (0.5mmol) of 1(R), 2(R)-DACH was dissolved in absolute ethanol in a separatebeaker, and the second solution was then added dropwise with gentlestirring to the first solution. After a clear solution was obtained, thesolution was maintained under refrigeration, and after a few days,yellow crystals were collected and used for single crystal X-raydiffraction and elemental analysis (Table 4).

1(R),2(R)-[(DACH)₂Au]Cl₃ (2) was synthesized by directly mixing oneequivalent of Na[AuCl₄].2H₂O with two equivalents of diamine ligand inalcoholic media using a previously described method (S. S. Al-Jaroudi,M. Monim-ul-Mehboob, M. Altaf, M. Fettouhi, M. I. M. Wazeer, S.Altuwaijri, and A. A. Isab, New J. Chem., vol. 38, no. 7, pp. 3199-3211,2014, incorporated herein by reference in its entirety). First, 200 mg(0.5 mmol) of NaAuCl₄.2H₂O was dissolved in absolute ethanol, 114 mg(1.0 mmol) of 1(R),2(R)-DACH was dissolved in absolute ethanol in aseparate beaker, and the second solution was then added dropwise withgentle stirring to the first solution. The white crystals that formedwere collected, washed, and used for single crystal X-ray diffractionand elemental analysis (Table 4).

TABLE 1 Elemental data and melting points for the synthesized complexesMelting Found (Calc.)% Complex point (° C.) H C N (1) 172 decomp. 4.02(3.97)  16.4 (15.88) 6.47 (6.17) (2) 176 decomp. 5.24 (5.96) 24.75(23.85) 9.57 (9.27)

Example 3 Solution NMR Measurements

All NMR measurements were performed using a JEOL JNM-LA 500 NMRspectrometer. The ¹³C NMR resonance spectra were obtained with ¹Hbroadband decoupling at a frequency of 125.65 MHz, and the peaks werereferenced relative to TMS. The spectral conditions were 32 k datapoints, 0.963 s acquisition time, 1.00 s pulse delay, and a 45° pulseangle. The proton NMR spectra were obtained at a frequency of 500.00MHz. The ¹H and ¹³C NMR chemical shifts of free ligand and bothcomplexes are given in Tables 1 and 2.

TABLE 2 ¹H NMR for the synthesized gold(III) complexes and free(1R,2R)-(−)-1,2-diaminocyclohexane in D₂O solvent ¹H (δ in ppm) H3,H6H4,H5 Compound H1,H2 (eq) H3,H6 (ax) (eq) H4,H5 (ax) 1(R),2(R)- 2.19m1.69m 1.54m 1.13m 0.99m DACH (1) 2.97m 2.07m 1.54m 1.36m 1.04m (2) 3.02m2.12m 1.55m 1.47m 1.09m m: multiplet

TABLE 3 ¹³C NMR for the synthesized gold(III) complexes and free(1R,2R)-(−)-1,2-diaminocyclohexane in D₂O solvent ¹³C (δ in ppm)Compound C1,C2 C3,C6 C4,C5 1(R),2(R)-DACH 56.8 34.1 25.4 (1) 65.6 33.023.9 (2) 64.3 32.8 23.9

For both complexes 1 and 2, Tables 1 and 2 clearly show that theobserved number of ¹H and ¹³C resonances was half of the expectednumber, which might indicate the presence of a C₂ center of symmetry.All cyclohexyl protons showed a downfield shift relative to the freediaminocyclohexane ligand. At room temperature, the cyclohexyl ringbehaves as a rigid conformer that allows axial and equatorial protons inthe cyclohexyl rings in 1 and 2 to be distinguished upon complexationcompared with the free diaminocyclohexane. These observed shifts can beexplained by the donation of nitrogen lone pairs to the d orbitals ofthe gold(III) center. This donation caused the nitrogen atoms to be moreelectron-deficient and made the H1 and H2 protons of the cyclohexyl ringmore deshielded, thereby shifting their resonances downfield, indicatingthat coordination took place through the nitrogen atoms. The resonancesfor other protons were shifted downfield for the same reason. Theobserved resonances for the complexes 1 and 2 are different. Thisvariation is due to their different geometries.

Similar downfield shifts were observed for the ¹³C resonances of C1 andC2 adjacent to their amino groups. These two carbons become deshieldeddue to coordination through nitrogen atoms, and the other carbons areshifted upfield.

Example 4 Solid-State NMR Measurements

Solid-state ¹³C NMR spectra were recorded at 100.613 MHz on a Bruker 400MHz spectrometer at an ambient temperature of 298 K. Samples were packedinto 4-mm zirconium oxide (ZrO) rotors. Cross-polarization andhigh-power decoupling were employed. A pulse delay of 7.0 s and acontact time of 5.0 ms were used in the CPMAS experiments. The magicangle spinning (MAS) rates were maintained at 4 kHz. Carbon chemicalshifts were referenced to tetramethylsilane (TMS) by setting thehigh-frequency isotropic peak of solid adamantane to 38.56 ppm. Thesolid-state NMR data is given in Table 3.

TABLE 4 Solid-state NMR for synthesized gold(III) complexes CompoundC1,C2 C3,C6 C4,C5 (1) 68.5 36.9 26.5 (2) 66.9 36.4 26.3

In the solid state ¹³C NMR spectra, significant deshielding was observed(as shown in FIG. 1) for complex 2. A similar trend was observed in thechemical shifts of complex 1, shown in Table 3. This observation showsthat the complexes are stable in solution as well as in the solid state.

Example 5 IR and Far-IR Spectroscopy

A PerkinElmer FTIR 180 spectrophotometer was used to collect IR spectrafor the ligands and synthesized gold(III) complexes within the range of4000-400 cm⁻¹. Far-infrared spectra were recorded for the synthesizedcomplexes 1 and 2 at room temperature with 4 cm⁻¹ resolution usingpolyethylene disks with a far-IR beam splitter.

Both complexes 1 and 2 showed significant N—H stretching vibrations,with an observed blueshift relative to the free ligand within the rangeof 3200-3500 cm⁻¹. This is due to coordination through an N atom and aformation of a five-membered ring with gold(III), which reduces Hbonding compared with free amino groups in free DACH.

The far-IR spectrum of complex 1 shows a clear vibration band at 367cm⁻¹, which is assigned to Au—Cl stretching vibration, with another bandat 393 cm⁻¹ for the Au—N bond (S. S. Al-Jaroudi, M. Fettouhi, M. I. M.Wazeer, A. A. Isab and S. Altuwaijri, Polyhedron, vol. 50, pp. 434-442,2013, incorporated herein by reference in its entirety). Complex 2lacked symmetric Cl—Au—C; stretching vibrations at 352 and 367 cm⁻¹.

Example 6 X-Ray Crystallography

Suitable crystals of complexes 1 and 2 were obtained as pale yellowblocks and colorless rods, respectively, from absolute ethanol. Theintensity data of the complexes were collected at 173K (−100° C.) on aStoe Mark II-Image Plate Diffraction System equipped with a two-circlegoniometer and using Mo-Kα graphite monochromated radiation (λ=0.71073Å) (Stoe & Cie. (2009). X-Area & X-RED32. Stoe & Cie GmbH, Darmstadt,Germany, incorporated herein by reference in its entirety). Thestructure was solved by direct methods with SHELXS-2014 (G. M.Sheldrick, Acta Cryst. A64, pp. 112-122, 2008, incorporated herein byreference in its entirety). The refinement and all further calculationswere carried out with SHELXL-2014. The H atoms from water were locatedon difference Fourier maps and refined using the distance restraint ofO—H=0.84(2) Å and with U_(iso)(H)=1.5 U_(eq)(O). It was not possible tolocate both H atoms on water molecules, O7W and O8W, for complex 2.

The N- and C-bound H-atoms were included in the calculated positions andtreated as riding atoms: N—H=0.91 Å and C—H=0.99-1.00 Å withU_(iso)(H)=1.2 U_(eq)(N,C). The non-H atoms were refined anisotropicallyusing weighted full-matrix least-squares on F². A semi-empiricalabsorption correction was applied using the MULscanABS routine in PLATON(A. L. Spek, Acta Cryst. D65, pp. 148-155, 2009, incorporated herein byreference in its entirety). FIGS. 3-7 showing the crystal structures andcrystal packing were drawn using Mercury software (C. F. Macrae, I. J.Bruno, J. A. Chisholm, P. R. Edgington, P. McCabe, E. Pidcock, L.Rodriguez-Monge, R. Taylor, J. van de Streek and P. A. Wood, J. Appl.Cryst. 41, pp. 466-470, (2008), incorporated herein by reference in itsentirety). The summary of crystallographic data of complexes 1 and 2 isgiven in Table 6.

TABLE 6 Crystal data and structure refinement details for complexes 1and 2 Complex 1 2 CCDC deposit no. 1055595 1055495 Chemical formula2[C₆H₁₄N₂AuCl₂]⁺•2Cl⁻•H₂O [(C₆H₁₄N₂)₂Au]³⁺•3(Cl)⁻•4(H₂O) Molecularweight 853.03 603.76 Crystal system, space group Monoclinic, P2₁Monoclinic, P2₁ Temperature (K) 173 173 a, b, c (Å) 9.5692 (5), 8.5645(5), 7.4766 (3), 27.3859 (10), 14.3950 (8) 10.7744 (5) β (°) 95.369 (4)90.913 (4) V (Å³) 1174.57 (11) 2205.81 (16) Z 2 4 Radiation type Mo KαMo Kα μ (mm−1) 13.17 7.06 Crystal size (mm) 0.40 × 0.38 × 0.35 0.45 ×0.35 × 0.30 Diffractometer STOE IPDS 2 STOE IPDS 2 diffractometerdiffractometer Absorption correction Multi-scan Multi-scan (MULscanABSin (MULscanABS in PLATON; PLATON; Spek, 2009) Spek, 2009) T_(min),T_(max) 0.251, 1.000 0.448, 1.000 No. of measured, independent 16303,4442, 4214 30420, 8309, 7854 and observed [I > 2σ(I)] reflectionsR_(int) 0.142 0.079 (sin θ/λ)_(max) (Å−1) 0.609 0.609 R[F2 > 2σ(F2)],wR(F2), S 0.045, 0.117, 1.01 0.021, 0.043, 0.94 No. of reflections 44428309 No. of parameters 179 470 No. of restraints 1 21 Largest diff. peakand hole (e Å⁻³) 1.65, −2.38 1.08, −1.27 Δρ_(max), Δρ_(min) (e Å⁻³)Absolute structural parameter 1.08, −1.27 −0.018 (4)

The crystal structure of complex 1 is shown in FIG. 3. The compoundcrystallized with two [(C₆H₁₄N₂)AuCl₂]+ cations and two Cl⁻ anions in anasymmetric unit, together with a water molecule for which it was notpossible to locate the H atoms. The gold(III) ion was bonded to the twonitrogen atoms of the (1R,2R)-(−)-1,2-diaminocyclohexane ligand and twochloride ions in a distorted square planar geometry. The Au—N bonddistances in molecule 1 and molecule 2 of complex 1 were in the range of2.020(15) to 2.059(16) A, respectively, while the Au—Cl bond distanceswere in the range of 2.261(5) to 2.281(5) A, respectively, as shown inTable 7. The Cl—Au—Cl and N—Au—N bond angles were 91.4(2) to 93.71(17)and 83.2(6) to 83.7(6) ° for molecules 1 and 2, respectively. The Au—Nbond distances in molecule 1 were significantly different from eachother, whereas the Au—N bond distances in molecule 2 were very close toeach other but different from those of molecule 1. The N—Au—N bond anglevalues reflected the chelation strain of the diamine ligand. Thesevalues were similar to those found in gold(III) complexes anddichloro-(trans-(±)-1,2-diaminocyclohexane)-gold(III) chloride hydrates(M. Monim-ul-Mehboob, M. Altaf, M. Fettouhi, A. A. Isab, M. I. M.Wazeer, M. N. Shaikh and S. Altuwaijri, Polyhedron, vol. 61, pp.225-234, 2013; and S. S. Al-Jaroudi, M. Fettouhi, M. I. M. Wazeer, A. A.Isab and S. Altuwaijri, Polyhedron, vol. 50, pp. 434-442, 2013, eachincorporated herein by reference in their entirety). The cyclohexyl ringadopted a chair conformation. There was a square planar geometry aroundthe Au(III) ion, resulting in a five-membered ring formation. All theamine groups were engaged in hydrogen bonding with water molecules andthe Cl⁻ counter ions. A water molecule was present in the crystallattice. The metal complex molecules packed head to head to generatemolecular chains along the c-axis, which in turn packed into layersparallel to the ac-plane (FIG. 4).

The X-ray structure of complex 2 is shown in FIG. 5. The compoundcrystallized with two [(C₆H₁₄N₂)₂Au]³⁺ cations, six Cl⁻ anions, andeight water molecules in an asymmetric unit. It was not possible tolocate both H atoms on water molecules O7W and O8W. In both molecules ofcomplex 2, the metal ion bonded to the four nitrogen atoms of two(1R,2R)-(+)-1,2-diaminocyclohexane ligands in a distorted square planargeometry. The Au—N bond distances were in the range 2.021(8)-2.048(6) A,and the N—Au—N chelate bond angles were in the range of 83.1(2)-83.6(2)° for molecules 1 and 2, as given in Table 7. These values were similarto those reported for [Au{trans-(±)-(1,2-DACH)}₂]Cl₃ andbis(ethylene-1,2-diamine)-gold(III) tris(perrhenate) (E. V. Makotchenkoand I. A. Baidina, J. Struct. Chem., vol. 52, pp. 572-576, 2011,incorporated herein by reference in its entirety). The cyclohexyl ringsadopted a chair conformation. The square planar geometry and thefive-membered ring strain imposed torsion angles for N1-C1-C2-N2 of54.0(8) ° and for N3-C7-C8-N4 of 58.3(8) ° for molecule 1, and forN5-C13-C14-N6 of 52.4(8) ° and for N7-C19-C20-N8 of 50.6(8) ° formolecule 2. The amine group hydrogen atoms were involved in hydrogenbonding interactions with Cl⁻ counter ions and water molecules,generating a three-dimensional hydrogen bonding network, as shown inFIGS. 6 and 7.

TABLE 7 Selected bond lengths (Å) and bond angles (°) for complexes 1and 2 Bond Lengths (Å) Bond Angles (°) Complex 1 Molecule 1 Au1—Cl12.269(6) Cl1—Au1—Cl2  91.4(2) Au1—Cl2 2.261(5) Cl1—Au1—N1 176.4(5)Au1—N1 2.020(15) Cl1—Au1—N2  92.8(5) Au1—N2 2.059(16) Cl2—Au1—N1 92.2(5) Cl2—Au1—N2 175.6(5) N1—Au1—N2  83.7(6) Molecule 2 Au2—Cl32.281(5) Cl3—Au2—Cl4  93.71(17) Au2—Cl4 2.275(5) Cl3—Au2—N3 173.6(5)Au2—N3 2.041(16) Cl3—Au2—N4  92.6(5) Au2—N4 2.028(15) Cl4—Au2—N3 90.5(5) Cl4—Au2—N4 175.8(4) N3—Au2—N4  83.2(6) Complex 2 Molecule 1Au1—N1 2.039(6) N1—Au1—N2  83.1(2) Au1—N2 2.044(6) N1—Au1—N3 179.2(2)Au1—N3 2.041(6) N1—Au1—N4  95.6(2) Au1—N4 2.044(6) N2—Au1—N3  97.7(2)N2—Au1—N4 178.2(3) N3—Au1—N4  83.6(2) Molecule 2 Au2—N7 2.048(6)N7—Au2—N8  83.2(2) Au2—N8 2.037(6) N5—Au2—N8 177.4(3) Au2—N5 2.021(8)N5—Au2—N6  82.8(2) Au2—N6 2.046(6) N5—Au2—N7  97.6(2) N6—Au2—N7 178.0(3)N6—Au2—N8  96.5(2)

Example 7 Interaction of Gold(III) Complexes with Glutathione (GSH) andUV-Vis and Kinetics Measurements

All kinetics measurements were made using a UV-vis spectrophotometer(Agilent Technologies, Cary 100 UV-Vis). Scanning kinetic mode was usedto investigate the change in the electronic spectra for gold(III)complexes upon interaction with GSH to determine the effectivewavelength for the reactions. Then, the kinetic mode was used to monitorthe reaction progress at the selected wavelength. A solution containing0.2 mM gold(III) complexes was prepared in water, and 40 mM sodiumchloride was added to improve the stability of the mononuclear-gold(III)complexes by preventing hydrolysis of the chlorides (A. Djeković, B.Petrović, Z. D. Bugar{hacek over (c)}ić, R. Puchta, and R. van Eldik,Dalton Trans., vol. 41, no. 13, pp. 3633-41, 2012, incorporated hereinby reference in its entirety). A series of GSH solutions was alsoprepared. Equal volumes of gold(III) complex solution and GSH solutionwere mixed, and the spectral changes recorded using the scanning kineticmode in the range from 200 nm to 350 nm. Then, the kinetic mode was usedto follow the reaction at three different temperatures. The reactionbetween the gold(III)-diamine complexes and the GSH ligand to formgold(III)-complexes can be illustrated in following equation:

The rate (k) of the described reactions can be calculated from the slopeof the linear plot of k_(obs) versus GSH concentration according to eqn(1), where the rate of the reverse reaction (k⁻¹) is obtained from theintercept of the straight line.

k _(obs) =k[GSH]+k ⁻¹ [Cl]  Eqn (1)

Using Microsoft Excel, the rate constants and activation parameters werecalculated from Eyring plots:

$\begin{matrix}{{\ln \; \frac{k}{T}} = {\frac{{- \Delta}\; H1}{RT} + {\ln \left( \frac{kB}{h} \right)} + \frac{\Delta \; S\; }{R}}} & {{Eqn}\mspace{14mu} (2)}\end{matrix}$

The kinetics of the substitution reaction between the synthesizedanticancer gold(III) complexes 1 and 2 with tripeptide glutathione wereinvestigated spectrophotometrically by monitoring the spectral changesof the gold(III) complexes in solution with excess of GSH. A blueshiftwas observed in all tested compounds, and the appearance of anabsorption band at 297 nm indicated the formation of a substitutionproduct with the formula [Au(DACH)(SG)₂]⁺. Equal volumes of thegold(III) complexes and GSH were kept until thermal equilibration wasachieved and then mixed directly inside a UV cell. The changes in theabsorption at a selected wavelength (297 nm) under pseudo-first orderconditions as functions of the glutathione concentration and temperaturewere collected. The pseudo-first order rate constant (k) was obtainedfrom the linear plot of k_(obs) versus the total GSH concentration (1).NaCl was added in excess to avoid the hydrolysis ofmononuclear-gold(III) diamine chloride, but this was not found to beeffective for complex 2 (FIG. 2).

From the data given in Table 5, the values of the rate constants andactivation parameters (ΔH‡, ΔS‡ and Ea) indicate that complex 1 reactsthree times faster than complex 2 in a substitution reaction with GSHdue to several factors related to the environment around the gold(III)center. This difference in reactivity can be attributed to ease of thechloride replacement by the GSH sulfur atom in complex 1 to form theshort lifetime intermediate [Au(DACH)(SG)₂]⁺ at 297 nm. However, theDACH ligand geometry also played a very important role in the reactivityof the gold(III) center, as the chair conformation of DACH in itsgold(III) complexes increased the stability by hindering GSH attack.Thus, the rate constant increased by a factor of 3. The decrease in theelectrophilicity of the metal center due to the electronic inductiveeffects of cyclohexyl in complex 2 also contributed to a slower reactionrate. The negative entropy of activation may be rationalized in terms ofthe formation of the intermediate, where two bulky GSH molecules aretied to the complexes, thereby reducing the entropy through the decreasein the number of reacting species.

TABLE 5 Kinetic data for the reaction of synthesized gold(III) complexeswith GSH ^(a)k ΔH^(‡) ΔS^(‡) Ea Complex (M⁻¹ · s⁻¹) (kJ · mol⁻¹)(JK⁻¹mol⁻¹) (kJ · mol⁻¹) (1) 114 × 10⁻² 30 −137 32 (2)  31 × 10⁻² 17−189 19 ^(a)Pseudo-first order rate constant at 298K.

Example 8 In Vitro Cytotoxic Assay for Synthesized Gold(III) Complexes

Complexes 1 and 2 were dissolved (10 mM) in DMSO and stored at −80° C.at volumes of 500 μL or aliquots with volumes of 10 μL were taken andstored at −20° C. (used once without refreezing). The compounds werethen diluted in RPMI medium immediately before use. Cisplatin waspurchased from Mayne Pharma. Culture medium with the same amount ofdrug-free DMSO was used as a negative control.

The classical Hodgkin lymphoma L-540 cell line was obtained from theGerman Collection of Microorganisms and Cell Cultures (Braunschweig,Germany). Human ovarian epithelial carcinoma-derived cancer cells A2780and its cisplatin-resistant clone A2780cis were from Sigma, Inc. (St.Louis, Mo., USA). The parent cisplatin-resistant subclone A2780cis wasmaintained by weekly treatment with 1 μM cisplatin (N. Casagrande, M.Celegato, C. Borghese, M. Mongiat, A. Colombatti, and D. Aldinucci,Clin. Cancer Res. vol. 20, pp. 5496-5506, 2014, incorporated herein byreference in its entirety). Cells were cultured at 37° C. in 5% CO₂ inRPMI medium supplemented with 10% heat-inactivated FBS, 0.1% (w/v)L-glutamine and antibiotics (0.2 mg/mL penicillin and streptomycin).

For the cell proliferation assay, 4.0×10³ A2780 and A2780cis cells wereseeded in 96-well flat-bottomed microplates in RPMI medium (100 μL) andincubated at 37° C. in a 5% CO₂ atmosphere for 24 h (to allow celladhesion) before drug testing. The medium was then removed and replacedwith fresh medium containing the compounds to be tested and cisplatin atincreasing concentrations (from 1 to 100 μM) at 37° C. for 72 h. Eachtreatment was performed in triplicate. Cell proliferation was assayedusing the MTT assay. L-540 cells (2.0×10⁵/ml) were seeded in 48-wellplates and treated with increasing concentrations of complex 1 andcomplex 2. After 72 hours, the number of viable cells was evaluated by atrypan blue dye exclusion.

The IC₅₀ values (i.e., the half maximal inhibitory concentration,representing the concentration of a substance required for 50%inhibition in vitro) were calculated using CalcuSyn software (Biosoft,Ferguson, Mo., USA) (T. C. Chou, and P. Talalay, Adv. Enzyme Regul. vol.22, pp. 27-55, 1984, incorporated herein by reference in its entirety).The results were presented as the mean±SEM of three replicate wells fromthree independent experiments. The cells were imaged under an invertedmicroscope (Eclipse TS/100, Nikon) with a photomicrographic system (DSCamera Control Unit DS-L2). Phase contrast micrographs were taken at amagnification of 4× (for L-540 cells) and 10× (for A2780 and A2780ciscells).

Statistical analysis was performed on the data using GraphPad Software.The significance of the differences was determined by Student's t-testfor comparison between two groups. P<0.05 was considered statisticallysignificant. The symbol “*” in FIGS. 8J, 8K, 9S, and 9T denotes P<0.05for gold complex versus control medium.

The in vitro antiproliferative activity of complexes 1 and 2 in theclassical Hodgkin lymphoma-derived cell line L-540 was evaluated (FIGS.8A-8K). Exposure of L-540 cells to increasing concentrations of bothgold complexes resulted in a dose-dependent cytotoxic activity, as shownin representative micrographs (FIGS. 8B-8I). Complex 2 exhibited a morepotent inhibition of cell growth compared to complex 1 (FIGS. 8J and8K), as evaluated by the trypan blue assay. Complex 2 was 7-fold moreactive than complex 1, with IC₅₀ values of 9.8 μM and 70.4 μM,respectively.

The cytotoxic activities of gold complexes 1 and 2 were also evaluatedin the ovarian carcinoma-derived cell line A2780 and in itscisplatin-resistant clone (A2780cis) using the MTT assay. For comparisonpurposes, the cisplatin activity was evaluated under the sameexperimental conditions. Both compounds inhibited proliferation in adose-dependent manner (FIGS. 9B-9I and 9K-9R). Again, complex 2exhibited a more potent inhibition of cell growth compared with complex1 in both A2780 (FIG. 9S) and A2780cis cells (FIG. 9T).

The IC₅₀ values of complex 1 were 61.3 μM for A2780cis and 15.7 μM forA2780 cells, and for complex 2, the IC₅₀ values were 31.1 μM forA2780cis and 6.98 μM for A2780 (Table 8). The IC₅₀ values for cisplatinin A2780cis and A2780 were 17.6 and 1.8 μM, respectively, showing a9.7-increase in resistance (fold resistance) to cisplatin in A2780ciscompared with A2780 (Table 8). Although the IC₅₀ values for both goldcomplexes were higher than those for cisplatin in A2780 and A2780ciscells, the fold-resistances of both complexes were approximatelytwo-fold lower than that of cisplatin (Table 8).

TABLE 8 Growth inhibition by complexes 1 and 2 in A2780 and A2780cisovarian cancer cells Fold resistance IC₅₀ (μM) A2780cis/A2780 ComplexA2780 A2780cis ratio Cisplatin 1.8 ± 0.1 17.6 ± 1.5 9.77 (1) 15.7 ± 1.4 61.3 ± 5.5 3.90 (2) 6.9 ± 0.6 31.1 ± 2.8 4.45

Altogether, these results indicate the higher cytotoxic potential ofcompound 2 with respect to compound 1 and the decreased fold-resistanceof both compounds with respect to cisplatin.

The foregoing discussion discloses and describes merely exemplaryembodiments of the present disclosure. As will be understood by thoseskilled in the art, the present disclosure may be embodied in otherspecific forms without departing from the spirit or essentialcharacteristics thereof. Accordingly, the disclosure of the presentdisclosure is intended to be illustrative, but not limiting of the scopeof the disclosure, as well as other claims. The disclosure, includingany readily discernible variants of the teachings herein, defines, inpart, the scope of the foregoing claim terminology such that noinventive subject matter is dedicated to the public.

1: A method for treating cancer, comprising: administering an effectiveamount of at least one of a gold(III) complex represented by formula(I), a gold(III) complex represented by formula (II):

a pharmaceutically acceptable salt, solvate, prodrug, or a combinationthereof to a subject; wherein the cancer is ovarian cancer, Hodgkinlymphoma, or both; each of R₁ is independently selected from the groupconsisting of a hydrogen and a methyl group; each of R₂ is independentlyselected from the group consisting of a hydrogen, a halogen, a hydroxyl,an amino, a nitro, a cyano, an optionally substituted alkyl, anoptionally substituted cycloalkyl, an optionally substitutedheterocyclyl, an optionally substituted arylalkyl, an optionallysubstituted heteroaryl, an optionally substituted alkoxyl, an optionallysubstituted aryl, an optionally substituted alkenyl, a N-monosubstitutedamino group, and a N,N-disubstituted amino group; and X is a chlorideanion.
 2. (canceled) 3: The method of claim 1, wherein R₂ is hydrogen.4. (canceled) 5: The method of claim 1, wherein the gold(III) complexrepresented by formula (I) is

and the gold(III) complex represented by formula (II) is

6: The method of claim 1, wherein the gold(III) complex represented byformula (I) and/or the gold(III) complex represented by formula (II) isadministered to the subject as the pharmaceutically acceptable saltcomprising at least one counterion which is at least onepharmaceutically acceptable anion selected from the group consisting offluoride, chloride, bromide, iodide, nitrate, sulfate, phosphate,methanesulfonate, ethanesulfonate, p-toluenesulfonate, salicylate,malate, maleate, succinate, tartrate, citrate, acetate, perchlorate,trifluoromethanesulfonate, acetylacetonate, hexafluorophosphate, andhexafluoroacetylacetonate. 7: The method of claim 6, wherein the atleast one counterion is chloride. 8: The method of claim 1, wherein thecancer is ovarian cancer, and the ovarian cancer is resistant tocisplatin. 9: The method of claim 1, wherein the cancer is Hodgkinlymphoma and the Hodgkin lymphoma is classical Hodgkin lymphoma. 10: Themethod of claim 1, further comprising measuring a concentration of abiomarker and/or detecting a mutation in the biomarker before and/orafter the administering. 11: The method of claim 10, wherein thebiomarker is at least one selected from the group consisting of BRCA1,BRCA2, CCL17, CD163, CD30, NF-κB, Gal-1, CA125, HE4, mesothelin,transthyretin, ApoA1, VCAM, IL-6, IL-8, B7-H4, serum amyloid A,transferrin, osteopontin, kallikreins, OVX1, VEGF, AGR-2, inhibin,M-CSF, uPAR, EGF receptor, lysophosphatidyl acid, beta2-microglobulin,miRNA, and Epstein-Barr virus DNA. 12: The method of claim 11, whereinthe concentration of the biomarker is measured with an ELISA assayand/or the mutation in the biomarker is measured with a PCR assay. 13:The method of claim 1, wherein the subject is a mammal. 14: The methodof claim 1, wherein the effective amount of the at least one of thegold(III) complex represented by formula (I), the gold(III) complexrepresented by formula (II), the pharmaceutically acceptable salt,solvate, prodrug, and a combination thereof is in a range of 1-100mg/kg. 15-20. (canceled)